Objective To explore the osteogenic potential of cervical intervertebral disc fibroblasts in vitro, to investigate the regulatory factors of recombinant human bone morphogenetic protein 2(rhBMP-2) and tumor necrosis factor α(TNF-α) on osteogenic phenotype of fibroblasts and to discuss the condition that facilitates osteogenesis of fibroblasts. Methods Theannulus fibroblasts cell lines of experiment goats were established in vitro and the biologicspecificity was found. According to different medias, 4 groups were included in this experiment: control group, TNF-α group ( 50 U/ml TNF-α), rhBMP-2 group (0.1 μg/ml rhBMP-2) and TNF-α+rhBMP-2 group (50 U/ml TNF-α+0.1 μg/ml rhBMP-2). Thefibroblasts were incubated in the media for about 3 weeks,and then the markers for osteogenic features were investigated by biochemistry, histochemistry observations. Results rhBMP-2 and TNF-α had no effect on the proliferation of fibroblasts from the experiment goats. rhBMP-2 or TNF-α could stimulate fibroblasts to secrete alkaline phosphatase and collagen type Ⅰ. The combined use of rhBMP-2 and TNF-α or the single use of rhBMP-2 could make fibroblasts to secrete osteocalin and the morphological changes of the fibroblasts were very obvious. Histochemical study of the nodules with specific new bone labeler(Alizarin red S) revealed positive reaction, denoting that the nodules produced by the fibroblasts werebone tissues. There was statistically significant difference(Plt;0.05) inALP activity between 3 experimental groups and control group and in secretion of osteocalcin between rhBMP-2 group, TNF-α+rhBMP-2 group and control group. Conclusion The results point out clearly that rhBMP-2 can induce theosteogenic potential of annulus fibroblasts in vitro.
On the basis of established JF305 cell line from human pancreatic cancer at this university, cell clone technique, cell electrophoresis, flower cytometer, and cancer orthotopically implanted nude mice technique were used to establish the sublines with different metastatic potential from human pancreatic cancer line-JF305 and the nude mice model implanted orthotopically with human pancreatic cancer monoclonal sublines with different metastatic potential. The results showed that the monoclonal cell sublines with different metastatic potential from human pancreatic caner-JF305 and the nude mice model implanted orthotopically with the sublines, would provided a useful method to study the metastatic mechanism of human pancreatic cancer.
OBJECTIVE: To investigate the characteristics and the pathologic classification of electrical-injury nerve using somatosensory evoked potential(SEP) technique. METHODS: SEP were detected and evaluated in 12 cases with electrical-injury nerve during operation, electrical stimulation was commenced from distal side of nerve where the structure of nerve looks normal under operating microscope, up to proximal side until evoking out a stable SEP predeterminate virtual value. Pathological examination and the following functional evaluation were compared with the values of SEP. RESULTS: At the site of nerve looking normal under operating microscope, perineurium appears normal or slightly thicken. But there are obvious fibrosis and fibrotic proliferation between fascicular and intrafascicular. Vessel plexus is not seen. At SEP stabilizely evoked site, nervous construction is normal, there are visible interfascicular vessel plexus and connective tissue appears loose. Comparing SEP values with pathological section, amplitude and latency of SEP is positively correlative with the quality of nerve. Eight cases repaired with SEP technique to select the anastomosis site for nerve transplantation were followed up, two-point discrimination reached grade III (America hand surgery association criterion) within 62.5% cases. CONCLUSION: SEP technique is valuable method for functional evaluation of electrical- injury nerve which has a complicated pathology. The pathology of electrical-injury nerve can be classified into 4 types, type A: fibrosis of nerve; type B: nerve looking normal under operation microscope, perineurium appears thicken, and there are obvious fibrosis and fibrotic proliferation between fascicular and intrafascicular, vessel plexus is rarely to see; type C: nerve looks normal, lymphocyte infiltration exists and it is obvious that there are many physalis-like, retrogressive construction in the section; type D: nervous construction is normal, there are visible interfascicular vessel plexus, and connective tissue appears loose, SEP always can be stably evoked.
High-frequency steady-state asymmetric visual evoked potential (SSaVEP) provides a new paradigm for designing comfortable and practical brain-computer interface (BCI) systems. However, due to the weak amplitude and strong noise of high-frequency signals, it is of great significance to study how to enhance their signal features. In this study, a 30 Hz high-frequency visual stimulus was used, and the peripheral visual field was equally divided into eight annular sectors. Eight kinds of annular sector pairs were selected based on the mapping relationship of visual space onto the primary visual cortex (V1), and three phases (in-phase[0o, 0o], anti-phase [0o, 180o], and anti-phase [180o, 0o]) were designed for each annular sector pair to explore response intensity and signal-to-noise ratio under phase modulation. A total of 8 healthy subjects were recruited in the experiment. The results showed that three annular sector pairs exhibited significant differences in SSaVEP features under phase modulation at 30 Hz high-frequency stimulation. And the spatial feature analysis showed that the two types of features of the annular sector pair in the lower visual field were significantly higher than those in the upper visual field. This study further used the filter bank and ensemble task-related component analysis to calculate the classification accuracy of annular sector pairs under three-phase modulations, and the average accuracy was up to 91.5%, which proved that the phase-modulated SSaVEP features could be used to encode high- frequency SSaVEP. In summary, the results of this study provide new ideas for enhancing the features of high-frequency SSaVEP signals and expanding the instruction set of the traditional steady state visual evoked potential paradigm.
As a noninvasive neuromodulation technique, transcranial magnetic stimulation (TMS) is widely used in the clinical treatment of neurological and psychiatric diseases, but the mechanism of its action is still unclear. The purpose of this paper is to investigate the effects of different frequencies of magnetic stimulation (MS) on neuronal excitability and voltage-gated potassium channels in the in vitro brain slices from the electrophysiological perspective of neurons. The experiment was divided into stimulus groups and control group, and acute isolated mice brain slices were applied to MS with the same intensity (0.3 T) at different frequencies (20 Hz and 0.5 Hz, 500 pulses) respectively in the stimulus groups. The whole-cell patch clamp technique was used to record the resting membrane potential (RMP), action potential (AP), voltage-gated potassium channels current of hippocampal dentate gyrus (DG) granule cells. The results showed that 20 Hz MS significantly increased the number of APs released and the maximum slope of a single AP, reduced the threshold of AP, half width and time to AP peak amplitude, and improved the excitability of hippocampal neurons. The peak currents of potassium channels were decreased, the inactivation curve of transient outward potassium channels shifted to the left significantly, and the time constant of recovery after inactivation increased significantly. 0.5 Hz MS significantly inhibited neuronal excitability and increased the peak currents of potassium channels, but the dynamic characteristics of potassium channels had little change. The results suggest that the dynamic characteristics of voltage-gated potassium channels and the excitability of hippocampal DG granule neurons may be one of the potential mechanisms of neuromodulation by MS.
Objective To investigate the relationship between graded spinal cord ischemia/reperfusion injury and somatosensory evoked potentials(SEP),neurologic function score（NFS）and the histopathological changes of spinal cord. Methods Forty rabbits were randomized and equally divided into 4 groups: shamoperation group, ischemia for 30min, 45min and 60min groups. The spinal cord ischemiareperfusion injury model was created by occlusion of the abdominal aorta in rabbits. SEP was monitored before ischemia,5,10minutes after ischemia, 15, 30 minutes, 1,2, 24 and 48 hours after reperfusion. NFS was evaluated at 6,12,24 and 48 hours after reperfusion.The pathological changes of spinal cord were observed after reperfusion 48 hours. Results The pathological characters with mild,moderate and severe spinal cord ischemia/reperfusion injury could be simulated by declamping after 30， 45 and 60 minutes infrarenal aorta crossclamping. SEP amplitude returned to normal after reperfusion 15 minutes（Pgt;0.05）and SEP latency returned to normal after reperfusion 30 minutes（Pgt;0.05）during mild spinal cord ischemia/reperfusion injury.SEP amplitude returned to normal after reperfusion 30 minutes（Pgt;0.05）and SEP latency returned to normal after reperfusion 60 minutes（Pgt;0.05）during moderate spinal cord ischemia/reperfusion injury. SEP latency increased and SEP amplitude decreased during severe spinal cord ischemia/reperfusion injury,compared with other groups, there were significant differences in SEP latency and SEP amplitude by clamping the infrarenal aorta for 60min（Plt;0.01）. With graded spinal cord ischemia/reperfusion injury, compared with shamoperation group, spinal cord ischemiareperfusion groups had significant differences in NFS（Plt;0.01）. Conclusion SEP is much quicker in the recovery of amplitude than latency during spinal cord ischemia/reperfusion. SEP is a sensitive and accurate index for spinal cord function during ischemia/reperfusion injury. SEP monitoring spinal cord ischemia/reperfusion injury during operation provides experimental basis for clinical application.
OBJECTIVE: To explore the pluripotential and possible clinical application of adult stem cells. METHODS: The original articles on adult stem cells were extensively reviewed and the recent advances were summed up. RESULTS: Adult stem cells were located at different tissues of human beings and had the pluripotentiality of self-renewal and differentiation. Some adult stem cells, such as in marrow, nerve, muscle, fat, skin, liver, tissues, had the ability to differentiate into the unrelated cell type. CONCLUSION: The pluripotential, ubiquitous distribution and plasticity of adult stem cells offered a new way in regeneration medicine, such as cell therapy and tissue engineering.
Motor imaging therapy is of great significance to the rehabilitation of patients with stroke or motor dysfunction, but there are few studies on lower limb motor imagination. When electrical stimulation is applied to the posterior tibial nerve of the ankle, the steady-state somatosensory evoked potentials (SSSEP) can be induced at the electrical stimulation frequency. In order to better realize the classification of lower extremity motor imagination, improve the classification effect, and enrich the instruction set of lower extremity motor imagination, this paper designs two experimental paradigms: Motor imaging (MI) paradigm and Hybrid paradigm. The Hybrid paradigm contains electrical stimulation assistance. Ten healthy college students were recruited to complete the unilateral movement imagination task of left and right foot in two paradigms. Through time-frequency analysis and classification accuracy analysis, it is found that compared with MI paradigm, Hybrid paradigm could get obvious SSSEP and ERD features. The average classification accuracy of subjects in the Hybrid paradigm was 78.61%, which was obviously higher than the MI paradigm. It proves that electrical stimulation has a positive role in promoting the classification training of lower limb motor imagination.
Neuron is the basic unit of the biological neural system. The Hodgkin-Huxley (HH) model is one of the most realistic neuron models on the electrophysiological characteristic description of neuron. Hardware implementation of neuron could provide new research ideas to clinical treatment of spinal cord injury, bionics and artificial intelligence. Based on the HH model neuron and the DSP Builder technology, in the present study, a single HH model neuron hardware implementation was completed in Field Programmable Gate Array (FPGA). The neuron implemented in FPGA was stimulated by different types of current, the action potential response characteristics were analyzed, and the correlation coefficient between numerical simulation result and hardware implementation result were calculated. The results showed that neuronal action potential response of FPGA was highly consistent with numerical simulation result. This work lays the foundation for hardware implementation of neural network.
Low-intensity ultrasound stimulation of the retina has the ability to modulate neural activity in the primary visual cortex (V1), however, it is currently unclear how different intensities and durations of ultrasonic stimulation of the retina modulate neural activity in V1. In this paper, we recorded local field potential (LFP) signals in the V1 brain region of mice under different ultrasound intensities and different stimulation times. The amplitude of LFP corresponding to 1 s before ultrasound stimulation to 2 s after stimulation (–1–2 s) was analyzed, including the power and sample entropy of delta, theta, alpha beta, and low gamma frequency bands. The experimental results showed that, as the stimulation intensity increased, the peak value of the LFP in the visual cortex showed a linear upward trend; the power in the delta and theta frequency bands showed a linear upward trend, and the sample entropy showed a linear downward trend. With increases of stimulation duration, the peak value of the LFP in the visual cortex showed an upward trend, and the upward trend gradually weakened; the power in the delta frequency band showed an upward trend, the sample entropy showed a linear upward trend, and the sample entropy in the theta frequency band showed a downward trend. The results show that low-intensity ultrasonic stimulation of the retina has a significant modulatory effect on neural activity in the visual cortex. The study provides insights into the mechanisms by which ultrasonic stimulation regulates visual system function. Furthermore, it clarifies the patterns of parameter selection, facilitating the development of personalized multi-parameter modulation for the treatment of visual neural degeneration, retinal disorders and related research areas.